Antirestriction Protein ArdB (R64) Interacts with DNA

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Antirestriction Protein ArdB (R64) Interacts with DNA

Biochemistry. Biokhimiia. 2020.

Манухов И. В., Кудрявцева А. А.

Abstract—The antirestriction ArdB protein inhibits the endonuclease activity of type I restriction/modification (RM) systems in vivo; however, the mechanism of inhibition remains unknown. In this study, we showed that recombinant ArdB from Escherichia coli cells copurified with DNA. When overexpressed in E. coli cells, a portion of ArdB protein formed insoluble DNAfree aggregates. Only native ArdB, but not the ArdBΔD141 mutant lacking the antirestriction activity, copurified with DNA upon anionexchange and affinity chromatography or total DNA isolation from formaldehydetreated cells. These observations confirm the hypothesis that ArdB blocks DNA translocation via the R subunits of the R2M2S complex of type I RM enzymes

Language: English

DOI

Keywords: antirestriction

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The ardA genes are present in a wide variety of conjugative plasmids and play an important role in overcoming the restriction barrier. To date, there is no information on the chromosomal ardA genes. It is still unclear whether they keep their antirestriction activity and why bacterial chromosomes contain these genes. In the present study, we confirmed the antirestriction function of the ardA gene from the Bifidobacterium bifidum chromosome. Transcriptome ...

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ArdB, ArdA, and Ocr proteins inhibit the endonuclease activity of the type I restriction-modification enzymes (RMI). In this study, we evaluated the ability of ArdB, ArdA, and Ocr to inhibit different subtypes of Escherichia coli RMI systems (IA, IB, and IC) as well as two Bacillus licheniformis RMI systems. Furthermore we explored, the antirestriction activity of ArdA, ArdB, and ...

Added: May 23, 2023

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Манухов И. В., Кудрявцева А. А., FEMS Microbiology Letters 2018

Antirestriction proteins of the ArdB group (ArdB, KlcA) specifically inhibit restriction (endonuclease) activity of restriction-modification (RM) type I systems. Antirestriction activity of KlcA and ArdB, encoded in transmissible plasmids RP4 (IncPα) and R64 (IncI1), respectively, has been determined. We show that the protein KlcA (RP4), an amino acid sequence identical to that of the protein ...

Added: October 19, 2022

The Importance of C-Terminal Aspartic Acid Residue (D141) to the Antirestriction Activity of the ArdB (R64) Protein

Манухов И. В., Кудрявцева А. А., Molecular Biology 2017

Abstract⎯Antirestriction proteins of the ArdB/KlcA family are specific inhibitors of restriction (endonuclease) activity of type-I restriction/modification enzymes. The effect of conserved amino acid residues on the antirestriction activity of the ArdB protein encoded by the transmissible R64 (IncI1) plasmid has been investigated. An analysis of the amino acid sequences of ArdB homologues demonstrated the presence ...

Added: October 19, 2022